BCG (Bacilli Calmette Guerin), an avirulent strain of Mycobacterium bodies, is one of the most widely used human vaccines in the world. The vaccine is extremely safe. We have recently developed vectors and the methodology to introduce recombinant DNA molecules stably into BCG and have been able to express foreign genes in BCG for the first time. Two types of E. coli as a plasmid. Shuttle phasmids have been developed. The first was a novel vector, termed a shuttle phasmis, that replicates in mycobactria as a phage and in E. coli as a plasmid. Shuttle phasmids have been constructed using temperate mycobacterial chromosomes. These vectors have been successfully used to integrate into mycobacteial chromosomes. These vectors have been successfully used to introduce and stably express a gene encoding kanamycin-resistance, the first selectable marker gene for mycobacteia. We have subsequently used kanamycin- resistance as a selection to develop a second set of vectors, E. coli-mycobacteria shuttle plasmids, which are the first transformation system for mycobacteria, including BCG. We plan to use these vectors and genetic methodologies to develop BG as a recombinant mutivaccine vehicle. This type of vaccine vehicle has novel immunological properties as it is the most potent immunological adjuvant known, especially for engendering cell- mediated immunity. In this proposal, we plan to clone and express a variety of HIV-antigens in this vaccine vehicle including synthetic polypeptides containing HIV B cell, T-helper cell, and cytotoxic T cell epitopes obtained from Projects 1 and 3 of this NCVDG> These ecombinant vaccines will be assedded in mouse and guinea pig animal model systems for their ability to produce T-cell immunity, T-cell memory, and humoral components of the immune response to the HIV-antigens. This approach should lead to a novel vaccine for the prevention and therapy of AIDS.